1.Effects of FGF-2 on the intercellular communication in the cardiac myocytes
Kequn SONG ; Yawen ZHANG ; Shiwe WANG
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
Objective To investigate the effects of FGF 2 on the intercellular communication in the cardiac myocytes. Methods The primary neonatal ventricular myocytes of rats were cultured with FGF 2 (0,1,10,100 and 1000ng/ml) for 1 or 4 days. Then the mean fluorescence recovery rate (MFRR, %/min) of these myocytes labeled with carboxyfluocein diacetate after photo bleaching was examined with laser scanning cytopmetry (ACAS Ultima). Results The MFRR levels of the myocytes cultured with FGF 2 (10 and 100ng/ml) groups for 1 day were significantly decreased compared with the control group, while no statistic changes were found among the other groups (FGF 2, 1 and 1000ng/ml) and the control. After FGF 2 exertion on the myocytes for 4 days, the increased cell number, enlarged cellular body and decreased MFRR level were found in the myocytes cultured with FGF 2 (10 and 100ng/ml) groups. And the decreased cell number, disarranged and shrunk cells and significantly increased MFRR level were gotten in the group (FGF 2 1000ng/ml) compared with the control group. Conclusion These results suggested the FGF 2 may have both short and long term effects on the cell cell communication in the cardiac myocytes mediated by gap junction, and be necessary to the cardiac myocyte coordination during the cell growth, differentiation and perhaps apoptosis.
2.Down-regulation of Beclin1 inhibits the resistance of ovarian cancer A2780/DDP cells for the cisplatin
GUO Yunhong ; TIAN Xiaoyu ; DENG Qiaozi ; ZHANG Kequn
Chinese Journal of Cancer Biotherapy 2020;27(6):615-621
[Abstract] Objective: To investigate the effect of Beclin1 knockdown on cisplatin resistance in ovarian cancer A2780 cells and its related mechanisms. Methods: The mRNA and protein expressions of Beclin1 in A2780 cells and drug resistant A2780/DDP cells were determined by qPCR and Western blotting. After transfection with Beclin1 siRNA, the sensitivity of A2780/DDP cells to cisplatin was detected by MTT assay; Cell clone formation and apoptosis were detected by the Colony formation assay and Flow cytometry assay, respectively; cell autophagy was monitored by monodansylcadaverin (MDC) staining. Furthermore, the protein levels of cell autophagy related proteins, lysosomal associated membrane protein Lamp-2 and Cathepsin B were detected by Western blotting. Results: The mRNA and protein expression levels of Beclin1 in cisplatin-resistant A2780/DDP cells were significantly higher than those in A2780 cells (all P<0.05). The expression of Beclin1 was significantly increased in A2780 cells after treated with cisplatin (P<0.05). Beclin1 knockdown promoted cisplatin induced apoptosis of A2780/DDP cells (P<0.05), inhibited autophagy and cell colony formation (all P<0.05), and increased cell sensitivity to cisplatin (P<0.05). Meanwhile, Western blotting showed that Beclin1 knockdown increased the protein levels of cleaved-caspase 3 and Cathepsin B in A2780/DDP cells, while down-regulated the protein expressions of Atg3, Atg7, LC3Ⅱ/Ⅰand Lamp-2 (all P<0.05). Conclusion: Beclin1 knockdown can improve the sensitivity of A2780/DDP cells to cisplatin, and the mechanism may be related to the inhibition of protective autophagy of cells by regulating the expressions of autophagy related proteins, and the regulation of lysosomes, thus further promoting cisplatin-induced apoptosis of drug-resistant cells.